title: Conditional and specific cell ablation in the marine annelid Platynereis dumerilii.
abstract: The marine annelid Platynereis dumerilii has become a model system for evo-devo, neurobiology and marine biology. The functional assessment of its cell types, however, has so far been very limited. Here we report on the establishment of a generally applicable, cell type specific ablation technique to overcome this restriction. Using a transgenic strain expressing the bacterial enzyme nitroreductase (ntr) under the control of the worm's r-opsin1 locus, we show that the demarcated photoreceptor cells can be specifically ablated by the addition of the prodrug metronidazole (mtz). TUNEL staining indicates that ntr expressing cells undergo apoptotic cell death. As we used a transgenic strain co-expressing ntr with enhanced green fluorescent protein (egfp) coding sequence, we were able to validate the ablation of photoreceptors not only in fixed tissue, using r-opsin1 riboprobes, but also by monitoring eGFP+ cells in live animals. The specificity of the ablation was demonstrated by the normal presence of the eye pigment cells, as well as of neuronal markers expressed in other cells of the brain, such as phc2, tyrosine hydroxylase and brn1/2/4. Additional analyses of the position of DAPI stained nuclei, the brain's overall neuronal scaffold, as well as the positions and projections of serotonergic neurons further confirmed that mtz treatment did not induce general abnormalities in the worm's brain. As the prodrug is administered by adding it to the water, targeted ablation of specific cell types can be achieved throughout the life of the animal. We show that ablation conditions need to be adjusted to the size of the worms, likely due to differences in the penetration of the prodrug, and establish ablation conditions for worms containing 10 to 55 segments. Our results establish mtz/ntr mediated conditional cell ablation as a powerful functional tool in Platynereis.
authors.affiliation: Max F. Perutz Laboratories, University of Vienna, Vienna, Austria ; Research Platform "Marine Rhythms of Life," Vienna, Austria.
authors.forename: Vinoth Babu
authors.initials: VB
authors.lastname: Veedin-Rajan
authors.forename: Ruth M
authors.initials: RM
authors.lastname: Fischer
authors.forename: Florian
authors.initials: F
authors.lastname: Raible
authors.forename: Kristin
authors.initials: K
authors.lastname: Tessmar-Raible
mesh_terms: D055011:Ablation Techniques / Q000379:methods*; D000818:Animals; D030801:Animals, Genetically Modified / Q000502:physiology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008795:Metronidazole / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D009601:Nitroreductases / Q000378:metabolism; D010786:Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D011077:Polychaeta / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D011355:Prodrugs / Q000494:pharmacology
chemical_list: D011355:Prodrugs; C485184:enhanced green fluorescent protein; D008795:Metronidazole; D049452:Green Fluorescent Proteins; D009601:Nitroreductases